Research Of The Protection With Asipirin In Primary Cultured Alveolar Epithelial Type II Cell Of The Rat

Objective To observe the protection of Asipirin in primary cultured alveolar epithelial type II cell(AEC-Ⅱ) of rat, and to discuss the mechanism of its antioxidant. Methods Purified AEC-Ⅱfrom the rat,which were primary cultured about 40h were established the oxidative damage model by H2O2. The AEC-Ⅱwere randomly divided into five groups in the first stage of the experiment (group C, group S, group A1, group A2, group A3):group C (Normal group); group S (damage group, add H2O20.5mmol/L, after primary cultured about 40h); group A1 (aspirin about 50μmol/L), group A2 (aspirin about 100μmol/L), group A3 (aspirin about 200μmol/L).Inverted microscope to observe the structure changes of the AEC-Ⅱ.3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) to test the A490. The AEC-Ⅱwere divided randomly into four groups in the second stage of the experiment (group C, groupAl, groupA2, groupA3), group C (Normal group);group A1 (aspirin about 50μmol/L), group A2(aspirin about 100μmol/L), group A3(aspirin about 200μmol/L). Collected the samples at 20h,40h,60h. Use the immunohistochemistry to test the protein of Heme oxygenase-1 (HO-1) and PCR to test the HO-1 mRNA. Results (1) This method could collect the AEC-Ⅱabout 2-2.5×107cell/rat. The purity and viability were over 90%. (2) The group S showed bigger intercellular space,less adherent cell, cell volume reduction and obviously lower cell_survival rate than the group C. The groupAl-3 showed less intercellular space,more adherent cell and obviously lower survival rate than the group S(P<0.05). (3) The group Al-3 showed the high expression of HO-1 protein and mRNA in the AEC-Ⅱthan the group C(P<0.05). Conclusion (1) This method could collect enough AEC-Ⅱwith better purity and viability by trypsinization and immunization adherency. (2) Aspirin could protect the primary cultured AEC-Ⅱof rat from the Oxidative damage. (3) Aspirin could improve the HO-1 protein and mRNA in the primary cultured AEC-Ⅱof rat. Up regulation HO-1 maybe one of the antioxidative mechanism of aspirin in the primary cultured AEC-Ⅱof rat.